Stability comparison of three candidate internal reference genes in Chinese giant salamander (Andrias davidianus)
Stability comparison of three candidate internal reference genes in Chinese giant salamander (Andrias davidianus)
复制标题
中国大鲵三个候选内参基因的稳定性比较
DOI：
10.1111/are.14383
复制
发表时间：
2019-11-12
影响因子：
2
通讯作者：
Deng, Dun
中科院分区：
文献类型：
--
作者：
Qian, Jing;Deng, Dun
作为食药两用的中国大鳗，一直是中国重点养殖的对象。研究中国大蜥蜴的基因功能需要通过使用适当的参考基因进行准确的归一化。本研究采用定量聚合酶链式反应技术，研究了β-肌动蛋白、谷氨酸脱氢酶脱氢酶和细胞色素b三个候选参考基因在不同组织、不同发育阶段和不同挑战条件下的表达水平。用geNorm、NormFinder和BestKeeper软件对这三个参考基因的稳定性进行分析。结果表明，在两个发育阶段和四种组织中，GAPDH的表达比β-肌动蛋白和细胞色素b更稳定。与GAPDH和Cytb相比，β-肌动蛋白在内毒素和GSIV处理的大鳗脾中最稳定。因此，在不同组织和不同发育阶段，GAPDH基因是比较合适的参考基因。该β-肌动蛋白基因可作为内毒素和GSIV攻击下大鳗脾组织中的一个参考基因。本研究为GAPDH和β-肌动蛋白分别在不同组织、不同发育阶段和不同挑战条件下作为中国大鳗合适的参考基因提供了令人信服的信息。
The Chinese giant salamander (Andrias davidianus) as food and medicinal product has been an important aquaculture object in China. Study of gene function in the Chinese giant salamander requires accurate normalization though the use of appropriate reference genes. In this study, the expression levels of three candidate reference genes including beta-actin, GAPDH and cytb of different tissues, different developmental stages and different challenges in Chinese giant salamander were evaluated by qPCR. The stabilities of these three reference genes were analysed by geNorm, NormFinder and BestKeeper software. The results showed that the expression of GAPDH was more stable than that of beta-actin and cytb in four tissues and at two developmental stages of Chinese giant salamander. Compared with GAPDH and cytb, beta-actin was the most stable in spleen of Chinese giant salamander treated with LPS or GSIV. Therefore, the result showed that GAPDH was the suitable reference gene in different tissues and at different developmental stages of Chinese giant salamander. The beta-actin could be used as a reference gene in spleen of Chinese giant salamander challenged with LPS and GSIV. This study provides convincing information for the GAPDH and beta-actin as suitable reference gene in Chinese giant salamander of different tissues, different developmental stages and different challenges respectively.